Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A, B) C3 (A) and C3aR1 (B) mRNA levels in the parahippocampal gyrus of NCI (n=32), MCI (n=34), or AD (Dementia, n=160). Spearman’s correlation, ρ (C3)=0.36, p<2.6E-08; ρ (C3aR)=0.33, p<3.4E-07.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A)Representative GFAP and Iba1 immunostainings with DAPI in the hippocampus of 9 month-old WT, C3aR KO, PS19, and PS19/C3aR KO mice. Scale bar: 50 μM.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Representative Western blots of PHF1- and CP13-positive phospho-tau and total tau in PS19 and PS19/C3aR KO mouse brains at 9 months. γ-tubulin is used as a loading control.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: Western Blot

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Representative synaptophysin (Syp) and Syp/NeuN co-immunostaining in area CA3 of hippocampus of WT, C3aR KO, PS19, and PS19/C3aR KO mice at 9 months. Scale bar: 50 μM.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: Immunostaining

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Unsupervised clustering gene expression heatmap from RNAseq analysis of hippocampal tissues of 9 month-old WT, C3aR KO, PS19, and PS19/C3aR KO mice. 1726 DEGs were identified by comparing the PS19 vs WT group (1155 genes upregulated, red, and 571 downregulated, blue). n=4-5/genotype, adjusted p<0.01.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: Expressing

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A) Gene-set enrichment analyses (GSEA) of the microglia sensome gene list in PS19 vs WT (left, NES=2.81, FDR<0.001) and in PS19/C3aR KO vs PS19 (right, NES= −3.0, FDR<0.001) comparisons. NES: normalized enrichment score; FDR: false discovery rate.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques:

Journal: Neuron

Article Title: Complement C3aR inactivation attenuates tau pathology and reverses an immune network deregulated in tauopathy models and Alzheimer’s disease

doi: 10.1016/j.neuron.2018.10.031

Figure Lengend Snippet: (A, B) Total STAT3 (A) and Y705-phospho-STAT3 (B) levels in 9 month-old WT (n=10), C3aR KO (KO, n=10), PS19 (PS, n=7), and PS19/C3aR KO (PS/KO, n=7) mice measured by RPPA (mean ± s.e.m). The asterisks above PS19 are for PS19 vs WT comparison; those above PS19/C3aR KO are for PS19/C3aR KO vs PS19 comparison. Student｀s t-test. *p<0.05; **p<0.01.

Article Snippet: After washing in TBS, sections or cultured cells on coverslips were blocked with TBS-T (0.4% Triton X-100) containing 2% donkey serum for 30 min, and then incubated in primary antibody diluted in blocking solution overnight at 4°C (mouse anti-GFAP (Millipore, AB_94844); rabbit anti- Iba1 (Wako, AB_839504); mouse anti-NeuN (Millipore, AB_2298772), rabbit anti-synaptophysin (Abcam, AB_2286949); rat anti-C3 (Hycult, AB_10129042); rat anti-C3aR (Hycult, AB_10130173); rabbit anti-pSTAT3 (Tyr705) (CST, AB_2491009)).

Techniques: